Biomod/2012/Titech/Nano-Jugglers/Protocols: Difference between revisions
No edit summary |
No edit summary |
||
| Line 1: | Line 1: | ||
{{Titech/Nano-Jugglers/HEAD}} | {{Titech/Nano-Jugglers/HEAD}} | ||
__NOTOC__ | __NOTOC__ | ||
=Protocols= | |||
<h2>PAGE protocol</h2> | <h2>PAGE protocol</h2> | ||
# Make 20% polyacrylamide gel.<br> | |||
# Put into a microwave oven and liquefy urea for 15 seconds.<br> | |||
# Wait until the solution become the room temperature.<br> | |||
# Add 10% APS 150 ul and TEMED 4ul.<br> | |||
# Put the solution into the gel box.<br> | |||
# Insert comb and wait it becomes coagulate(about 30 minutes).<br> | |||
# Make another gel box and prepare double gel boxes.<br> | |||
# Set up a tub for electrophoresis.<br> | |||
# Set double gel boxes and pour 1x TBE running buffer into the tub (upper tub’s buffer is new running buffer, and lower tub’s buffer is recycle buffer).<br> | |||
# Incline the tub to remove bubbles under gels.<br> | |||
# Clear wells of the gel by pipette to remove unharden gel solution.<br> | |||
# Connect the tub to a power source.<br> | |||
# Pre-run at specified voltage (250V or 300V) for 20 minutes.<br> | |||
# Clear wells of the gel by pipette again.<br> | |||
# Load wells with sample solution 5ul.(sample 3ul + 2x BPB solution 3ul = sample solution 6ul)<br> | |||
# Run at specified voltage (250V or 300V) for specified time (50 minutes)<br> | |||
# Put the gels into a cyber gold solution taper.(SYBR-Gold solution: 10000x cyber gold 20ul + MilliQ water 200ml)<br> | |||
# Shake the taper by hand for 10 minutes.(using used cyber gold solution, shake for 15 or 20 minutes)<br> | |||
# Take out the gels.<br> | |||
# Spot blue light to observe the gel.<br> | |||
# Take pictures through an orange filter by camera.<br> | |||
<h2>Observation of the platinum particles</h2> | <h2>Observation of the platinum particles</h2> | ||
# Make a solvent of about 100μL in an Eppendorf tube.<br> | |||
# Put beads in another Eppendorf tube with a spatula.<br> | |||
# Remove the dust on the surface of silicon rubber with the like Scotch tape.<br> | |||
# Drill holes in the silicon rubber with a belt punch.<br> | |||
# Press the holed silicon rubber to the surface of a dish.<br> | |||
# Add the beads in the tube to the solvent.<br> | |||
# Shake the tube by voltex and set in the Ultrasound device.<br> | |||
# Inject the beads solution into the hole of silicon rubber.<br> | |||
# Set the dish to the observation units after down the objective lens to the bottom.<br> | |||
# Observe with the eyepiece.<br> | |||
Revision as of 22:20, 31 August 2012
<html> <head> <style type="text/css"> /* ====================
主に全体に関わるCSS
==================== */ body.mediawiki {
font-size: 14px;
background-image:url(http://openwetware.org/images/2/2d/TNJback.png); background-repeat: repeat; font-family: Calibri, Verdana, helvetica, sans-serif; }
h1 {
padding: 0px 20px 5px 20px;
font-size: 34px;
line-height: 40px;
font-weight: bold;
}
h2 {
padding: 20px 20px 5px 20px;
font-size: 25px;
color: #000000;
text-decoration: none;
font-weight: bold;
}
h2 a {
color: #eb8300;
}
h3 {
padding: 20px 20px 5px 20px;
font-size: 20px;
color: #000;
font-decoration: none;
font-weight: bold;
}
h1.firstHeading {
display: none;
}
p { text-align: justify; } a:link { color: #00a5ea; text-decoration: none } a:visited { color:#00a5ea; text-decoration: none } a:hover {
color: #eb8300;
text-decoration: none } a:active { color:#f29400; text-decoration: none } #content{ margin: 0px 0 0px 0; align: center; padding: 12px 12px 12px 12px; width: 946px; background-color: #ffff; border: 0; } #bodyContent{ width: 970px; margin: 150px auto 0 auto; align: center; background-color: #ffffff; } #globalWrapper{ margin: 20px 0 0 0; width:994px; background-color: #ffffff; margin-left: auto; margin-right: auto } #biomodlink{ position:absolute;top:0px; width:970px;height:20px; background-color: #ffffff; margin:3px auto 3px auto }
/* ====================
メニューの画像を変更できる部分 ==================== */
#header {
margin: 0 auto 0 auto;
position:absolute;
top:28px; width:968px;height:189px;
background-color: #FFFFFF; background-repeat: no-repeat;
background-image: url(http://openwetware.org/images/7/74/Title-LOGO6.jpg);
} /* ====================
以下、特殊なclassに適用される ==================== */
#navigation { position:absolute;
top:120px; width:970px; height:92px;
background-repeat: no-repeat;
color: #0000FF; }
/* ====================
ここからプルダウン周辺のデザイン ==================== */
- menu * {
margin: 0; padding: 0; }
- menu {
position:absolute;
font-family: calibri, verdana, sans-serif;
font-color: #000000;
background-color: transparent; float:left; display: block; }
- menu ul {
position:relative;
float: left; text-align:center; list-style: none; }
- menu li {
background-color:yellow;
position:relative;
float:left;
width: 136px;
font-size: 23px;
font-weight: bold;
border:ridge 0px #FFFFFF; }
- menu li.ach {
background-color:yellow;
position:relative;
float:left;
width: 162px;
font-size: 23px;
font-weight: bold;
border:ridge 0px #FFFFFF; }
- menu li.sup {
background-color:yellow;
position:relative;
float:left;
width: 109px;
font-size: 17px;
font-weight: bold;
border:ridge 0px #FFFFFF; }
- menu li.none {
background-color:yellow;
position:relative;
float:left;
width: 152px;
font-size: 17px;
font-weight: bold;
border:ridge 0px #FFFFFF; }
- menu a {
color: #000000; display:block; text-decoration: none; }
- menu a:hover {
background-color:gold; }
- column-one { display:none; width:0px;}
.container{background-color: #ffffff; margin-top:0px} .OWWNBcpCurrentDateFilled {display: none;}
- footer{position: center; width: 994px}
@media screen {
body { background: #F5F5F5 0 0 no-repeat; /* changed default background */ }
} </style> </head> <BODY> <div id="biomodlink"> <<a href="http://openwetware.org/wiki/Biomod">BIOMOD</a>|<a href="http://openwetware.org/wiki/Biomod/2012">2012</a>|Titech Nano-Jugglers </div> <div id="header"> <div id="navigation"> <div id="menu"> <ul> <li><a href="http://openwetware.org/wiki/Biomod/2012/Titech/Nano-Jugglers"><br>Home<br><br></a></li> <li><a href="http://openwetware.org/wiki/Biomod/2012/Titech/Nano-Jugglers/Team/Students"><br>Team<br><br></a></li> <li><a href="http://openwetware.org/wiki/Biomod/2012/Titech/Nano-Jugglers/Project"><br>Project<br><br></a></li> <li><a href="http://openwetware.org/wiki/Biomod/2012/Titech/Nano-Jugglers/Results">Results<br>&<br>Methods</a></font></li> <li class="ach"><a href="http://openwetware.org/wiki/Biomod/2012/Titech/Nano-Jugglers/Achievements"><br>Achievements<br><br></a> <li class="sup"><a href="http://openwetware.org/wiki/Biomod/2012/Titech/Nano-Jugglers/Protocols"><br>Suppl. Info.<br><br></a></li> <li class="none"><a href="http://openwetware.org/wiki/Biomod/2012/Titech/Nano-Jugglers/Acknowledgement"><br>Acknowledgements<br><br></a></li> </ul> </div> </div> </div> </BODY> </html>
Protocols
PAGE protocol
- Make 20% polyacrylamide gel.
- Put into a microwave oven and liquefy urea for 15 seconds.
- Wait until the solution become the room temperature.
- Add 10% APS 150 ul and TEMED 4ul.
- Put the solution into the gel box.
- Insert comb and wait it becomes coagulate(about 30 minutes).
- Make another gel box and prepare double gel boxes.
- Set up a tub for electrophoresis.
- Set double gel boxes and pour 1x TBE running buffer into the tub (upper tub’s buffer is new running buffer, and lower tub’s buffer is recycle buffer).
- Incline the tub to remove bubbles under gels.
- Clear wells of the gel by pipette to remove unharden gel solution.
- Connect the tub to a power source.
- Pre-run at specified voltage (250V or 300V) for 20 minutes.
- Clear wells of the gel by pipette again.
- Load wells with sample solution 5ul.(sample 3ul + 2x BPB solution 3ul = sample solution 6ul)
- Run at specified voltage (250V or 300V) for specified time (50 minutes)
- Put the gels into a cyber gold solution taper.(SYBR-Gold solution: 10000x cyber gold 20ul + MilliQ water 200ml)
- Shake the taper by hand for 10 minutes.(using used cyber gold solution, shake for 15 or 20 minutes)
- Take out the gels.
- Spot blue light to observe the gel.
- Take pictures through an orange filter by camera.
Observation of the platinum particles
- Make a solvent of about 100μL in an Eppendorf tube.
- Put beads in another Eppendorf tube with a spatula.
- Remove the dust on the surface of silicon rubber with the like Scotch tape.
- Drill holes in the silicon rubber with a belt punch.
- Press the holed silicon rubber to the surface of a dish.
- Add the beads in the tube to the solvent.
- Shake the tube by voltex and set in the Ultrasound device.
- Inject the beads solution into the hole of silicon rubber.
- Set the dish to the observation units after down the objective lens to the bottom.
- Observe with the eyepiece.
